This study was designed to determine the distribution patterns of nidogen-1 (N-1) and -2 (N-2) in the rat uterus, oviduct and ovary. For this purpose, 6 female Wistar-albino rats found in proestrous period were used. The tissue samples fixed in 10% neutral formalin were examined immunohistochemically. Sections were incubated with primary goat-polyclonal N-1 and -2 antibody. Both the N-1 and -2 immunoreacted strongly with the uterine luminal and gland epithelium, many cells in the uterine stroma, the oviduct epithelium, the follicular cells of the unilaminar primary follicles, interstitial cells, and atretic follicles. They were moderately expressed in the follicular epithelium of primary and secondary follicles. The immunostaining intensity of N-1 was higher than that of N-2 in theca interna cells of graafian follicles. N-1 and -2 immunostaining were weakly detected in some of the primordial follicles, and in the granulosa lutein cells of corpus luteum. Neither theca externa nor zona pellucida immunoreacted with N-1 and -2. In conclusion, the present study showed that the nidogen are synthesized by the uterine luminal and the endometrial gland epithelium, the oviduct epithelium and ovarian follicles in the rats.
The distribution pattern of monoaminergic neurons in the common marmoset (Callithrix jacchus) was determined on the basis of comparative morphology using immunohistochemical methods with specific laboratory-made antibodies to tyrosine hydroxylase (TH), aromatic L-amino acid decarboxylase (AADC), dopamine-β-hydroxylase, phenylethanolamine-N-methyltransferase and serotonin (5-HT). Dopaminergic (DA) neurons (A16-A8 groups), noradrenergic neurons (A7-A1 groups), and adrenergic neurons (C1 and C2 groups) were found. Serotonergic neurons were first found in the rostral part of the nucleus interpedunculalis where they were mainly distributed in the raphe nuclei (B9-B1 groups). However, no neurons of the B4 group were found, contrasting with previous reports on rats. Characteristic findings on the distribution of monoaminergic neurons in the brain of the common marmoset were as follows: DA neurons other than those included in A groups were observed in the basal magnocellular nucleus; TH-only-immunoreactive (Ir) neurons and AADC-only-Ir neurons were occasionally found intermingled with DA neurons in the substantia nigra; and TH-Ir neurons coexisting with 5-HT-Ir neurons were found in the rostralmost area of the dorsal raphe nucleus. These results clearly show that patterns of monoaminergic neuron distribution in the brain of the common marmoset are unique and differ from those observed in rodents.
To study the relationship between aging and levels of monoaminergic neurons, 2,4-diamino-6-hydroxypyrimidine (DAHP), an inhibitor of monoamine synthesis, was intraperitoneally administered to senescence-accelerated mouse-prone (SAMP8) mice. Time course of immunoreactive intensity for serotonergic (5-HT) neurons in the dorsal raphe nucleus, which were stained using laboratory-raised serotonin-specific antibody, was quantitatively evaluated using an image analysis system. Results showed that 5-HT neurons are not highly sensitive to a synthesis inhibitor common to both catecholaminergic and 5-HT neurons.
To investigate the properties of endocytosis in the marginal cells of stria vascularis (SV), we perfused cationized ferritin (CF), an endocytosis tracer, into the cochlear duct. After five minutes to two hours of endolymphatic perfusion, the tissues were fixed and the distribution of CF was observed by transmission electron microscopy. CF was endocytosed by the marginal cells. CF-loaded vesicles were represented mainly by coated vesicles. The time-course of endocytosis was as follows. After five minutes perfusion, CF was distributed over the surface of the apical membrane and CF-loaded vesicles were seen very close to the apical membrane. After 30 minutes perfusion, a number of CF-loaded small endosomes and CF-loaded large vacuolar endosomes were observed at the supranuclear region. After one hour and two hours perfusion, CF was transferred to the multivesicular body. After two hours perfusion, acid phosphatase (ACPase) activity, a marker of lysosomes, was investigated. CF was not transferred to lysosomes until after two hours, because ACPase activity was not seen in CF-loaded vesicles. In this paper, we discuss the basic endocytosis properties of marginal cells.
Different types of integrin alpha subunit were investigated immunohistochemically to determine the relationship between the cell adhesion molecule and the local morphology of the rat anterior pituitary using anterior pituitary tissues and primary cultured cells. Alpha 3 subunit was shown to be the main integrin subunit in the anterior pituitary. In vitro, immunoreactions of α3 subunit appeared mainly surrounding cell clusters. The distribution roughly overlapped with immunoreaction of laminin, while it was also observed where expression of laminin was negative. When dispersed cells were primary cultured for 2, 3 or 5 days, immunoreaction of α3 subunit was clearly detected in the vicinity of cell membrane throughout the period; while that of α5 subunit was not observed on day 2, it was observed as dots in cytoplasm on day 3 and more distinctly as lines in the vicinity of the cell membrane on day 5. Alpha 5 subunit was not detectable in vivo. These results indicate the possibility that integrin α3 subunit not only binds the cells to the basement membrane but also to other extracellular matrix or directly to adjacent cells in the anterior pituitary. In addition, integrin α5 subunit may supplement the function of α3 subunit under certain conditions.
The stapes footplate articulates the vestibular window (stapediovestibular joint; SV joint). The SV joint is known as crucial region of otosclerosis. To shed a light on pathogenesis of otosclerosis, we have examined the histochemical characteristics of the rat ossicular joint cartilage. Interestingly, glycoconjugate histochemistry revealed that the cartilage matrix components of the SV joint were significantly different from those of the other ossicular joints, i.e. incudomalleolar (IM) and incudostapedial (IS) joints. Alcian blue, pH 2.5 and high-iron diamine procedures vividly stained the cartilage matrices of the IM and IS joints, while those of the SV joint were faintly stained. Chondroitin sulfate immunohistochemistry revealed intense labeling of the cartilage of the IM and IS joints, while the immunolabeling of the SV joint was negligible. On the contrary, keratan sulfate immunolabeling was discernible on the cartilage matrices of the SV joint, but not on those of the IM and IS joints. Type II collagen immunolabeling revealed that the marginal region of the IM and IS joint cartilage matrices was devoid of the immunostaining, while the SV joint cartilage matrices were uniformly labeled with this antibody. In this study, we have succeeded in demonstrating the histochemical characteristics of the rat SV joint cartilage matrices of which is quite different from those of the other ossicular joints.