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The cells of the seminiferous epithelium of the rat testis are a rich source of microtubules and contain distinct microtubular structures such as the meiotic spindle and manchette. Microtubule diversity can be maintained by differential genetic expression of the multiple alpha- and beta-tubulin polypeptides or by tubulin monomer acetylation and detyrosination, post-translational modifications of alpha-tubulin. In the present analysis, antibodies that specifically recognize acetylated (antiacetylated), tyrosinated (anti-Tyr) and detyrosinated (anti-Glu) alpha-tubulins were employed to examine the distribution of post-translationally modified microtubules in the cells of the seminiferous epithelium. In the light microscope, a distinct pattern of staining for each antibody was detected using immunoperoxidase techniques on paraffin-embedded testicular sections. In the case of the anti-Glu antibody, a dense immunoperoxidase staining was detected in the cytoplasm of steps 4-7 spermatids. Thereafter, staining was noted over the area corresponding to the manchette of steps 8-15 spermatids, but not over their cytoplasm. The tails of spermatids were also reactive with this antibody. The anti-Tyr antibody was observed to be localized over the cytoplasm of Sertoli cells in their basal, supranuclear, and apical regions. A dense immunoperoxidase staining was also noted in the cytoplasm of pachytene spermatocytes, but it was negligible in the cytoplasm of spermatocytes undergoing their meiotic division; in these cells the centrioles and meiotic spindle were reactive. The spermatid's tails were also reactive. The antiacetylated antibody showed reactivity only over the tails of spermatids. With the electron microscope, a similar pattern of labeling was noted using immunogold labeling on Lowicryl K4M embedded testicular sections. The anti-Glu antibody heavily labeled microtubules of the manchette and the axoneme of tails of spermatids as well as microtubules of the proximal and distal centrioles and centriolar adjunct. The anti-Tyr antibody strongly labeled microtubules of Sertoli cells and the meiotic spindle and midbody of dividing spermatocytes. The anti-Tyr antibody also labeled the microtubules of the axoneme, centrioles, and centriolar adjunct of spermatids, but to a lesser degree than the anti-Glu antibodies; the manchette was faintly labeled. Of the three antibodies, the antiacetylated antibody showed the weakest labeling of microtubules of the centrioles, centriolar adjunct, and midbody, whereas those of the manchette and Sertoli cells were unreactive; the axoneme was moderately labeled.(ABSTRACT TRUNCATED AT 400 WORDS)
Anat Rec 1991 Jan
PMID:Differential post-translational modifications of microtubules in cells of the seminiferous epithelium of the rat: a light and electron microscope immunocytochemical study. 199 83

The distribution of glutamylated tubulin has been analyzed in mammalian testis using the specific mAb GT335 by immunoelectron microscopy and immunoblotting. In spermatozoa of various species, immunogold labeling showed the presence of glutamylated tubulin in all of the microtubules of axoneme and centrioles, whereas the microtubule network of the spermatid manchette was unlabeled. In earlier germ cells, centriole was the only microtubule structure to be labeled. A similar distribution was observed using the anti-acetylated tubulin antibody (6-11B-1), confirming previous results of Hermo et al. [Anat. Rec. 229:31-50, 1991]. However, among testicular somatic cells, microtubules of some Sertoli cell branches were not acetylated but glutamylated. 2-D PAGE of mouse and hamster sperm extracts showed a high level of alpha and beta-tubulin heterogeneity, comparable to that found in brain. Immunoblotting with GT335 revealed a large amount of glutamylated tubulin resolved into numerous alpha as well as beta-tubulin isoforms. This suggests that the major testis-specific tubulin isotypes (m alpha 3/7 and m beta 3) are also glutamylatable. These results show a subcellular sorting of posttranslationally modified tubulin isoforms in spermatids, glutamylation being associated with the most stable microtubule structures.
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PMID:Differential distribution of glutamylated tubulin during spermatogenesis in mammalian testis. 791 Jul 83

Callitrichid primates (tamarins, marmosets) have extreme variation in the vomeronasal organ (VNO), including ontogenetic differences in the neuroepithelium and vomeronasal duct (VND) patency at birth. Such differences render the timing and extent of VNO maturation debatable in callitrichids, but no studies have used neuron-specific immunohistochemical markers to address this question. The present study compared the number of VNO epithelial cells that express immunoreactivity to neuron-specific beta-tubulin III (BT), VNO length, and VNO cross-sectional area between two species of tamarins (Leontopithecus rosalia and Saguinus geoffroyi) that differed in perinatal VND patency. Neonatal lemurs and adult marmosets and bushbabies were also examined for a comparison to species previously shown to have a relatively large amount of VNO neuroepithelium and patent VNDs. The head of each specimen was serially sectioned in the coronal plane. Based on known rostrocaudal start/stop points of the VNO, selected unstained sections were used for BT protocols and area measurement at three percentiles (25th, 50th, 75th) in each specimen. Each section was photographed and enlarged for cell counts and measurement of cross-sectional epithelial area. In each specimen, the number of BT(+) cells in the VNO was counted at each percentile and expressed as a number per mm(2). Results indicated that lemur VNOs had a dense population of BT(+) cells at birth, but the VNO was more varied in the tamarin species. S. geoffroyi had few or no BT(+) cells in VNOs of neonates, which had fused VNDs, but had an increased BT(+) population by 1 and 2 months postnatal age, when the VND was patent. Of the species with patent VNDs at birth, neonatal L. rosalia had a denser population of BT(+) cells compared to S. geoffroyi, though not to the degree seen in neonatal lemurs or adult marmosets and bushbabies. These findings show that BT immunohistochemistry is a useful comparative method for the study of VNOs in subadult primates. Since the quantity of nonsensory VNO epithelium varies substantially between species, epithelial area measurements may be misleading, and BT(+) cell counts appeared to be the best quantitative method for comparing receptor neuron numbers among primates. It is suggested that the greater BT(+) cell population in L. rosalia at all subadult stages examined reveals an earlier maturation of the neuroepithelium compared to S. geoffroyi. Further investigation should consider whether this may relate to a comparatively brief subadult ontogeny and early onset of adult behaviors in L. rosalia compared to other tamarins studied to date.
Anat Rec A Discov Mol Cell Evol Biol 2004 May
PMID:Ontogenetic observations on the vomeronasal organ in two species of tamarins using neuron-specific beta-tubulin III. 1510 35

Vomeronasal organ (VNO) morphology varies markedly across primate taxa. Old World monkeys display no postnatal VNO. Humans and at least some apes retain a vestigial VNO during postnatal life, whereas the strepsirrhines and New World Monkeys present a morphologically well-defined VNO that, in many species, is presumed to function as an olfactory organ. Available microanatomical and behavioral studies suggest that VNO function in these species does not precisely duplicate that described in other mammalian taxa. The questions of which species retain a functional VNO and what functions they serve require inquiry along diverse lines but, to be functional, the vomeronasal epithelium must be neuronal and olfactory. We used immunohistochemistry to establish these criteria in six primate species. We compared the expression of two neuronal markers, neuron-specific beta-tubulin (BT) and protein gene product 9.5, and olfactory marker protein (OMP), a marker of mature olfactory sensory neurons, in paraffin-embedded VNO sections from two strepsirrhine and four haplorhine species, all of which retain morphologically well-defined VNOs during postnatal life. The infant Eulemur mongoz, adult Otolemur crassicaudatus, neonatal Leontopithicus rosalia, and adult Callithrix jacchus express all three proteins in their well-defined vomeronasal neuroepithelia. The infant Tarsius syrichta showed some BT and OMP immunoreactivity. We establish that two strepsirrhine species and at least some New World haplorhines have mature sensory neurons in the VNO. In contrast, at all ages examined, Saguinus geoffroyi VNO expresses these markers in only a few cells.
Anat Rec A Discov Mol Cell Evol Biol 2004 Nov
PMID:Expression of neuron-specific markers by the vomeronasal neuroepithelium in six species of primates. 1547 Jun 76