The role of pituitary growth hormone in foetuses during intrauterine life has been the subject of various controversy. In this report I presented firstly the results on the time of appearance of growth hormone, and its content in human foetal pituitary glands, and also its concentration in cord blood. Then I discussed the transplacental passage and finally reviewed the regulation of growth hormone secretion in foetuses and neonates. Human foetuses were collected at the time of artificial abortion and immediately washed with saline and kept in acetone. Growth hormone was extracted from acetone dried foetal heads or pituitary regions by homoginizing in saline and subjected to radioimmunoassay. 48 foetuses were investigated in total, their gestational age ranging from 6 to 20 weeks. None of the foetus at the 6th week in gestation contained measurable growth hormone. However, in the 7th week, 2 out of 7 foetus contained a few mμg of growth hormone and in the 8th week, 6 out of 8 contained 2 to 13.5 mμg. By the 9th week, growth hormone was detected in all of 3 foetuses in amounts of 6 to 30 mμg. Growth hormone content in foetal pituitary glands increased logarithmically thereafter. By the end of the 2nd month they contained up to 10 mμg and by the end of the 3rd month, 10 to more than a hundred. At the 4th month from a hundred to more than a thousand and by the end of 5th month, 40 μg of growth hormone were being stored in a pituitary gland. Growth hormone content per gram of wet weight of the foetus showed a great spurt between the 4th and 5th month of gestation as compared to the previous stage.
Growth hormone concentration in cord blood was determined in 7 foetuses. One foetus at the 10th week of gestation contained 18 mμg/ml of growth hormone in its cord blood. At the 16th week, cord blood contained 40-120 mμg/ml of growth hormone.
Transplacental passage has been denied in many protein hormones as was proved in case of growth hormone at the time of delivery. Judging from the high concentration of growth hormone in cord blood far beyond that observed in mothers, transplacental passage of growth hormone was reuld out in early stage of pregnancy as well.
Time of appearance or synthesis of other pituitary hormones were reviewed, they range from the 7-8th week for growth hormone to 21st week for FSH.
As for the regulation of foetal growth hormone secretion several reports were reviewed. Human neonates are known to respond to insulin induced hypoglycemia. (Cornblath et al. 1965). In simian foetuses and neonates within 24 hours after delivery, arginine infusion does not increase growth hormone concentration in blood as reported by Mintz et al.1969. Cornblath et al. reported in 1965, that glucose administration increased paradoxically the serum growth hormone level in human neonates, and that it is not until 15 days after delivery when glucose suppresses growth hormone concentration in blood, the ressponse usually seen in children and adults. Mintz also observed the same paradoxical rise of growth hormone in simian foetuses and neonates when glucose was administered.
It has been the prevailing view that growth hormone is not indispensable for the foetal intrauterine growth and many investigators support this concept. Still foetal pituitary glands store and probably synthesize growth hormone as early as in 7 weeks of gestation and cord blood containes high level of growth hormone from the early stage of pregnancy as was just reported above. It is therefore suggested that foetal growth hormone may be playing an important role for the foetal growth and metabolism than was thought before.

抄録全体を表示

I. Plasma TSH in pregnant women.
Plasma TSH concentrations in 38 pregnant women from 8 to 32 weeks' gestation ranged in the levels of normal subjects (≤5.0 μU/ml) determined by HTSH radioimmunoassay.
II. Human Chorionic Thyroid-Stimulating factor.
The partial purification and immunological characteriaztion of the human chorionic thyroid-stimulating factor was described. Bates' percolation method and CM-cellulose chromatography were applied to normal human placenta, and allows the detection of human chorionic thyroid-stimulating factor by Mckenzie's bioassy. Multiple doses of the Bates' fraction and. CM fraction resulted in curves not parallel with that obtained by human TSH standard. The biological activity of human chorionic thyroid-stimulating factor was not neutralized by the in vitro addition of anti-bovine TSH serum. On the other hand, no significant effects were observed when Bates' fraction and CM fraction were assayed by the HTSH radioimmunoassay.
III. TSH in the human fetal pituitaries.
TSH concentration in human fetal pituitaries was determined by radioimmunoassay and the McKenzie's bioassay. In 40 human fetal pituitaries from 8 to 32 weeks' gestation, TSH determined by radioimmunoassay was first detectable at 12 weeks' gestation, and the content increased progerssively with fetal age and reached about 1/10 of TSH content in adult pituitary glands at 32 weeks' gestation. TSH content per fetal body weight was calculated and found to be maximum at 12 to 17 weeks' gestation. On the other hand, TSH activity determined by the McKenzie's bioassay in 30 human fetal pituitaries from 8 to 32 weeks' gestation was first detectable at 14 weeks' gestation. The ratio of bioassay potency (B) /radioimmunoassay potency (I) was 6.65 (mean) (in control experiment, the ratio of B/I of 5 adult pituitaries was 4.5), when Human Thyrotropin Research Standard A was used as a standard in both assays.

抄録全体を表示

The fetal TSH was investigated qualitatively and histochemically using fetal pituitaries. These fetuses were obtained from women who had abortions but did not have any thyroid disease. The TSH activity could be detected at the 8th week of gestation and the value was 2.15 mU/mg. From the 9th to the 12th week of gestation the value was 2.93 mU/mg. and from the 13th to the 18th week of gestation, it had rapid increase and ranged from 4.4 mU/mg to 4.9 mU/mg. From the 19th to the 22nd week of gestation or the 6th month of gestation the value was 6.44 mU/mg. There was a rapid increase of the value at the 23rd week of gestation and at the 25th week it came up to 8.2 mU/mg. There was also a marked increase in the weight of the fetal pituitary gland. At the 8th month of gestation the value ranged from 9.6 mU/mg to 13.5 mU/mg and at the 10th month it ranged from 15.9 mU/mg with its maximum value. On the other hand, in the histological study the anterior pituitary lobe showed a funicular structure. Among the anterior-lobe cell, α-cell and γ-cell could be found but β-cell was not clear. There was no PAS or Gomori-positive cell at this time. Even at the 6th month of gestation, there was still much interstitial connective tissue and the coexisistence of pseudofollicles and reticular structure was found. But the differentiation of these three cells, namely α, β and γ cells became possible at the 6th month of gestation compared to the 5th month. The same thing would be found with CHP stain and there was also PAS-positive cell. The reticular structure became obscure at the 7th month of gestation and the cells became more easy to be differentiated. The size of the cells got large and there was also Gomori-positive cells. At the 8th month of gestation, the number of β-cell became more significant and its specific morphology was very clear. It seems that the differentiation of the auterior pituitary lobe was established at this time. There was no histological change in the anterior lobe at the 9th and the 10th month of gestation and only an increase in the number and the size of the each cell were found. From the above events we can conclude that the fetal thyroid gland had the influence from its own fetal pituitary gland since the 12th week of gestation and the pituitarythyroid system was already completed by the time of birth.

抄録全体を表示

人胎盤から大量のprogesteroneとestrogenが分泌されていることは, 以前からよく知られていた.しかし, 人胎盤には3β-ol dehydrogenase と aromatizing enzyme system は豊富に存在するが, その他のhydroxylase, 側鎖断裂酵素がみとめられないので, cholestero1からのprogesterone, estrogenの生合成が行なわれ難く, その性ホルモン生合成機構は不明瞭であつた.近年のDiczfalusyを頂点とする研究^{1) 2) 3) 4)} により, progesterone生合成のためにはpregnenolone, estrogen 生合成のためにはandrogen が主として胎児副腎から人胎盤に供給され, 胎盤の強い3β-oldehydrogenase, aromatizingenzymeの作用によりfeto-Placental unitという型でprogesterone, estrogen が生合成されていることが明らかにされた.
以上の様に, feto-placental unitにおける性ホルモンの生合成は, 主として人について研究され解明されたため, その生合成は非常にactiveであるとの考えが通念となつた.しかし, 妊娠後半に卵巣を摘出しても流産をおこさない人と, 妊娠全期を通じて妊娠維持のために卵巣が必要な家兎, ラット等では, feto-placental unit における性ホルモンの生合成はことなる筈である.したがつて著者らは, feto-placentalunitにおける性ホルモン生合成についてほとんど研究されていない家兎を選び研索をすすめ, その機構を明らかにし, 人のそれと対比させようと試みた.

抄録全体を表示

It has been well known that there are two pathways to estriol (E3) from dehydroepiandrosterone (DHEA) in the feto-placental compartment. In A-pathway, in which the aromatization of DHEA in the placenta is antecedent and the 16 a-hydroxylation of estrone (El) in the fetus is followed, E3 is mainly formed in the fetal liver. In B-pathway, in which the 16α-hydroxylation of DEHE in the fetus is antecedent and the aromatization of 16α-hydroxy-C 19 steroids in the placenta is followed, E3 is finally formed in the placenta. The B-pathway is subdivided into B-1 and B-2 pathways. In the B-1 pathway, 16α-hydroxy-DEHA is first formed in the fetal adrenal or liver, and then E3 is formed through 16α-hydroxyandrostenedione and 16α-hydroxy-El in the placenta. In the B-2 pathway, androstenetriol is first formed in the fetal liver, followed by the formation of E3 through 16-hydroxytestosterone in the placenta.
In an attempt ot clarify the main pathway to E3 from DHEA in the feto-placental unit, H³-DHEA and C¹⁴-androstenedione were administered into the umbilical blood in two cases of midpregnant women in which interruption of preganancy was performed by cesarean section for medicosocial reasons.
The radioactive metabolites in the placenta, umbilical blood, fetal liver and maternal urine obtained 15 minutes later were analyzed. In the case 1, the ³H/C¹⁴ ratios of E3 in the placenta, umbilical blood and fetal liver were 5.5, 9.5 and 1.7, respectively. The respective ratios of El were 3.0, 1.0 and 0.9; those of 16α-hydroxy-E1 were 1.6, 1.0 and 1.0, respectively. The The ³H/C¹⁴ ratios of 16α-hydroxytestosterone in the placenta and umbilical blood were 19.8 and ∞, respectively. The ³H/C¹⁴ ratios of various steroids in the case 2 were comparatively the same as those in the case 1. In the placenta and umbilical blood, the ³H/C¹⁴ ratios of E₃ were markedly higher than those of 16α-hydroxy-E1. In constrast, the ratio of E3 in the fetal liver was almost the same as that of 16α-hydroxy-El. Furthermore, the quantity of E3 in the placenta and umbilical blood is approximately 2 times that in the fetal liver.
From these data, one could draw the following conclusion;
1) In the feto-placental compartment, E3 might be formed in the placenta in large quantities and in the fetal liver in smaller quantities.
2) E3 in the placenta and umbilical blood might be mainly formed from DHEA via 16α-hydroxytestosterone (B-2 pathway).
3) E3 in the fetal liver might be mainly formed from DHEA via El and 16α-hydroxy-El (A-pathway).

抄録全体を表示

It has been thought that testicular androgen is essential for the sexual differentiation of the genital ducts and the external genitalia which takes place at the early period of fetal life. It has been suggested recently, however, that androgen might have another important role for the sexual differentiation which is induced in the hypothalamus at an early neonatal period. According to Harris the hypothalamus is undifferentiated sexually still at birth at least in the rat. Male rats with castration soon after birth show completely female type of behavior as well as female pattern of cyclical hypophyseal gonadotropin secretion after their maturation. Conversely female rats with the administration of testosterone at the neonatal period develop male type of sexual function afterwards. The influence of androgen on the neonatal hypothalamus which otherwise develop functionally according to the female pattern regardless of the sex is assumed to be responsible for its development to the male type.
In the human, however, there have been few data available for the evaluation of the significance of androgen especially in the newborn.
In the present paper deals with the human subjects as well as the rats. In anencephalic foetus the retardation of testicular growth became stignifican at the later half of gestation when HCG levels decrease. In the newborn babies plasma LH activities as estimated by HCG-radioimmunoassay were higher at least during a first few days after birth in comparison with that observed in normal man, although they showed gradual decrease with the days of postnatal life. The rats of their decrease was lower than that expected if the LH activities observed were derived from placental sourse alone, suggesting the additional secretion of hypophyseal LH in the fetus or the newborn itself.
Plasma testosterone levels in the fetus as measured by double isotope derivative technique were not different between sexes, although the male had slightly more androstenedione than the female. After birth, however, androgen levels in the male babies both became significantly higher transiently than that observed before birth. On the other hand, the female babies showed only rapid decrease in both steroids.
It would be possible to make several suppositions from the data described above. Firstly, the large amount of HCG appearing during the earliest trimester of gestation may contribute to the morphological sexual differentiation occurring in the fetus. Secondly, the fetal hypophysis would begin to secrete LH from itself during the late pregnancy to stimulate the fetal gonads in place of HCG to decrease at this stage. Thirdly, the characteristic rise of androgen concentrations in the plasma observed in the male neonate may have some relation with functional sexual differentiation in the hypothalamus as supposed in the rat.

抄録全体を表示

Quantitative and dynamic studies on biosynthesis and relaese of human placental proteins and HPL in vitro were performed in six cases of normal term placentas. Fresh placental tissue divided into small fragments was incubated in KRB buffer containing ³H- 1-leucine for up to 96-120 hours. Amount of proteins, HPL, ³H-proteins and ³H-HPL in tissue extract and medium was analyzed.
Tissue extract and medium were fractionated by gel filtration on Sephadex G-100. The distribution of radioactivity showed three major peaks. Appropriate fractions were precipitated by specific antisserum to HPL. The distribution of radioactive proteins precipitated showed two peaks ; mol. wt. of the radioactive species of the first peak was greater than 100,000, and that the second peak was about 20,000 which was determined to be the same as mol. wt. of HPL or ¹³¹I-PHL. Immunological properties and electrophoretic mobility of the smaller mol. wt. radioactive proteins precipitated by anti-HPL were examined, and it was concluded that this radioactive aspecies were identied with ³H-HPL. Although the nature of the larger mol. wt. radioactive proteins precipitated by anti-HPL was not determined, an altenative explanation was discussed.
The amount of proteins in tissue decreased with incubation time but was variable among different placentas; and the amount of ³H-proteins in tissue increased with time but was variable. During the first 24 hours HPL content in tissue decreased and therafter the content remained fairly constant. Relatively few counts of ³H-HPL in tissue were detected in all cases except one (placenta E) which showed significantly higher counts. The cumulative amount of proteins and ³H-proteins in mdeium increased progerssively and was fairly constant among five placentas. Only placenta E had relatively higher counts of ³H-proteins in tissue. The amount of HPL and ³H-HPL in medium increased with incubation time but varied widely in different placentas.
The percentage of 'H-HPL to 3H-proteins in medium was 5-10% (four placentas) which was significantly higher than that in tissue (1-2%). In two placentas (E and F) the percentage in medium was strikingly high suggesting the possible existence of a biochemically abnormal placenta. These data indicated that HPL was preferentially released into medium.
It was calculatde that about 80% and 90% of the total synthexised ³H-HPL was released into medium within the first 24 hours and 48 hours respectively. While only 22-28% of the total ³H-proteins was found in medium in this period. These data suggested that the rate of release of newly synthesized ³H-HPL was very rapid compared with that of ³H-proteins.
The specific activity of ³-HPL in medium was extremely higher than that in tissue, and the difference increased with longer incubation time. While the ratio of spedific activity of ³H-proteins in medium to tissue was almost unity after 24-48 hours. These results suggested that there might be a HPL pool in placental tissue. This stable HPL in tissue was not readily released as newly synthesized ³H-HPL.
Some possible relationship between HPL and fetus was discussed.
(These studies have already been pubilshed in Endocrinology 85 (6) : 1028 and 1037, 1969. by Suwa, S. and H. Friesen.)

抄録全体を表示

HPL was extracted by modified Friesen's method from term placenta and was purified by gel filtration, ion exchange cellulose chromatography and DEAE-Sephadex A-50 chromatography. The purified HPL developed a single band by 7% polyacrylamide disc electrophoresis. The immunological character of this HPL showed evidence of partial identity with human growth hormone but antigenicity was not identiacl with human chorionic gonadotropin. It showed no cross reaction with prolactin and human sera. As to physicochemical character, the carbohydrate content was not recognized by the orcinol sulfuric acid method. No free sulfhydryl group was detected in 8M urea or 2M guanidine-HCI. This indicates that the sulfhydryl groups of HPL are located within the molecule rather than on the surface, since they are only accessible when the molecule is unfolded.
The biological character was also studied for its lipolytic action on rat epididymal fat pad in vitro. Addition of epinephrine (10 μg per ml), caffeine (0.01 M per ml), cyclic 3'5' AMP (1 mg per ml) or simple HPL (30 μg per ml) to the incubation medium released nearly equal amounts of FFA. But combination of HPL (30 ?Eg per ml and 3 μg per ml) plus caffeine (0.01 M per ml) gave an enhanced release of FFA and activated hormone sensitive lipase (HSL) and lipoprotein lipase (LPL). When adrenergic blockers (Dibenamine 3 μg per ml and Inderal 30 ?Eg per ml) were added in combination with HPL, HSL and LPL activities were markedly lowed, but release of FFA was not largely influenced. When castrated male rats were previously injected with estrogen 1μg per 100 g body weight for 4 days or progesterone 1 mg per 100 g body weight for 4 days, the release of FFA was accelerated by HPL in progesterone sensitized fat pad in vitro but estrogen had no influence. Injection of reserpin (100 μg per 100 g) had no effect of the release of FFA. When pregnant rats were injected with HPL, the FFA, insulin and glucose was markedly elevated in the maternal blood, while FFA in fetal blood increased significantly and insulin and glucose remained the same. The above findings indicate that HPL acts on maternal tissue, activates adenyl cyclase and accelerates lipolysis via cyclic AMP. Thus it has an important influence on the metabolism of fat and carbohydrate in the fetoplacental-maternal unit. This is probably an important factor for fetal growth.

抄録全体を表示

ホルモンの作用臓器における代謝という観点から雌性ラットにおけるOxytocinの代謝を子宮ホモジネ_トを用いて検討したところOxytocin不活性化効力およびその様式に臓器特異性が認められた.すなわち, 1) Oxytocin投与により子宮に特異的に不活性化効力が増大すること, 2) 卵巣摘出によつてもOxytocin投与の場合と同様に増大し, これにEstradiolを投与すると不活性化効力は減少し正常値をとることを認めた.また¹³¹I-Oxytocinの子宮におけるuptakeは卵巣摘出により著明に減少しEstradiolおよびProgesterone投与により増加することなどを認めた.

抄録全体を表示

甲摘及びチロキシン投与ラットの酸素消費量に対する環境温度, ツボクラリン, ペントバルビタールの効果を検討した.甲腺状状態が高いほど, 低温による酸素消費量の増加は少なかつた.チロキシン投与により上昇した酸素消費量はツボクラリンにより変化を見なかつたが, ペントバルビタールにより低下を見た.ペントバルビタール投与時の酸素消費量は甲状腺状態により明らかに示した.

抄録全体を表示

甲摘及びチロキシン投与ラットの酸素消費量に対するジニトロフェノール及びペントバルビタールとジニトロフェノールの効果を検討した.甲状腺状態が高いほど, ジニトロフェノール感受性は高かつた.ペントバルビタール前処置は甲摘群において, ジニトロフェノール感受性に変化を見なかつたが, チロキシン投与群において, 明らかな感受性の低下を見た.

抄録全体を表示

正常妊娠, 双胎妊娠, 妊娠申毒症の胎盤を用いてprogesterone代謝, 特にその生合成 (invitro) の面より追究した.
progesterone concentrationは, 胎児の性別や, その胎盤の一卵性, 二卵性であるかにより差異を認めない.
胎盤のslice incubation実験により, 正常胎盤 (単胎, 双胎) におけるprogesterone synthesisはNADPH-generating system下で著明な増加を認めたが, 妊娠中毒症の場合は低値であつた.
radioactive precursorsを用いてprogesterone synthesisを検討した.

抄録全体を表示

HPLの免疫学的検索の第一歩として著者らが作製した抗HPL血清を用いて, HPL, HCG, HGH及びcrude HPLについて基礎的検討を加えた.HPLとHGHとの間に交叉反応が認められたが, HPLとHCG, 及びHCGとHGHとの間には交叉反応が認められなかつた.免疫電気泳動ではHPLの方がHGHより陽極側に泳動されていた.胎盤から抽出したcrude HPLには, 血清成分の混在はあるが, 抽出法としては満足すべきものと考えられる結果を得た.

抄録全体を表示

従来TRFの化学構造について不詳の点が多かつたがFolkersらの合成になるTripeptide (pyro9-lutamylhistidilproline Amide) のTRF作用を検討した.この合成TRFはマウス, ラットを使用したin vivoの検定, ラット下垂体によるin vitroの検定の両者で明らかなTRF作用を有する事を観察した.同時にin vivoでは合成TRF投与量とTSH放出反応との間に用量反応関係を有し, 投与後10分でピークを示すTSH放出の時間経過を示した.
Fotkersらの合成になるTripeptide (pro-glutamylhistidilproline amide) のTRF作用を観察しこの合成TRFがTSHの賦活作用をもたず, TSH様作用を有せず, 既報のように投与量とTSHの反応との間に用量反応関係を有しin vivoとin vitroでの効果の一致性からみてそのTRF作用は特異的なものである事を証明した.合成TRFはT₄によつて抑制を受けこのT₄の抑制作用はT₄投与量と合成TRF投与量とに容量依存的である.

抄録全体を表示