Characterization of gap junctions between pairs of Leydig cells from mouse testis

Pérez-Armendáriz, E. Martha; Romano, Maria; Luna, J.; Miranda, Carolline Billett; Bennett, Michael V. L.; Moreno, Alonso P.

doi:10.1152/ajpcell.1994.267.2.c570

Cited by 61 publications

(52 citation statements)

References 24 publications

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“…The intimate anatomical association enables quick and specific access and action of their neuropeptides on Leydig cells. The gap junctions between Leydig cells further make themselves able to respond promptly to the neurotransmitters as a group (Pérez-Armendariz et al 1994). Due to lack of monoaminergic innervation, these peptidergic nerves may serve as major neuronal regulators for Leydig cell steroidogenesis in young adult men.…”

Section: Discussionmentioning

confidence: 99%

Peptidergic not monoaminergic fibers profusely innervate the young adult human testis

Gong

Feng

et al. 2009

Journal of Anatomy

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Numerous studies have reported that intratesticular nerves exert important regulatory effects on the functions of the male gonad; however, as yet little is known about their distribution in the young adult human testis. The purpose of this study was to explore whether peptidergic and adrenergic nerves occur in the male gonad of this age, and, if present, to depict their distribution further. Thirty testes were collected from 15 reproductively healthy donors aged 21-32 years. Antibodies against protein gene product 9.5 (PGP 9.5), neuropeptide Y (NPY), C-terminal flanking peptide of NPY (CPON) and vasoactive intestinal peptide (VIP) were employed for immunohistochemical detection of intratesticular peptidergic nerves, and those against dopamine-beta-hydroxylase (DBH) and 5-hydroxytryptamine (5-HT) for monoaminergic ones. The testicular parenchyma exhibited a rich innervation by PGP 9.5-positive fibers, mainly associated with Leydig cell nests, blood vessels, and seminiferous tubules. Numerous NPY-and CPON-immunoreactive (IR) nerves also appeared in the gonads, but the vast majority were confined to blood vessels. A small number of VIP-IR fibers were detected in some arterioles. By contrast, however, no fibers displaying DBH or 5-HT immunoreactivity were observed within the testis. Additionally, expression of PGP-9.5, NPY, CPON, VIP, DBH and 5-HT was found in Leydig cells, PGP 9.5 in spermatogonia, and NPY and CPON in peritubular myoid cells. Our results suggest that the young adult human testis is devoid of monoaminergic nerves but profusely innervated by peptidergic fibers, which may serve as major neuronal regulators for testicular functions at this age.

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Section: Discussionmentioning

confidence: 99%

Peptidergic not monoaminergic fibers profusely innervate the young adult human testis

Gong

Feng

et al. 2009

Journal of Anatomy

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“…Cryosections of 10 -12 m were processed as previously described (Pérez-Armendariz et al, 1994). Briefly, sections were fixed in 70% ethanol at Ϫ20°C; rinsed with phosphate buffer saline (PBS), pH 7.3, 1 mM CaCl 2 , and preincubated in 2% albumin IgG-free, 0.1% Triton X-100 in PBS.…”

Section: Immunocytochemical Localization Of Cx43 3␤hsd Mih and Gcnamentioning

confidence: 99%

“…A specific spatial distribution has been found for Cx26, Cx31, Cx32, Cx33 and Cx43 using IF studies in testis from adult animals (Batias et al, 1999;Mok et al, 1999;Pérez-Armendariz et al, 1994;Pelletier, 1996;Risley et al, 1992;Tan et al, 1996). Several lines of evidences support the idea that Cx43 is required for adequate control of fertility.…”

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confidence: 95%

“…Leydig cells have gap junctions (reviewed, Russell, 1996) and are extensively dye-coupled, both in vivo (Risley et al, 1992) and in freshly dispersed preparations (Kawa, 1987;Pérez-Armendariz et al, 1994;Varanda and de Carvalho, 1994). Cx43 is the only connexin described in the interstitium so far.…”

mentioning

confidence: 99%

“…Cx43 is the only connexin described in the interstitium so far. It has been localized in peritubular cells and in rodent (Batias et al, 1999;Pérez Armendariz et al, 1994;Pelletier, 1996;Risley et al, 1992) and human (Steger et al, 1999) Leydig cells. In mouse, functional studies support the idea that Cx43 is the main connexin communicating between adult Leydig cells: a) biophysical properties of macroscopic and microscopic currents recorded from double whole cell voltage clamp of isolated pairs of LC are similar to those recorded from deficiently connected cell pairs transfected with Cx43 (Pérez- Armendariz et al, 1994); b) decrease in Cx43 expression in LC clumps cultured for 36 hr occurs associated with an 80% decay in dye coupling (Pérez-Armendariz et al, 1996); and c) time-dependent decay in Cx43 and dye coupling are prevented by maintaining LC cultures in the presence of luteinizing (LH) or chorionic gonadotrophic (CG) hormones or dibutyryl-cAMP (Pérez-Armendariz et al, 1996).…”

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Expression of Connexin43 in mouse Leydig, Sertoli, and germinal cells at different stages of postnatal development

et al. 2001

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Connexin 43 (Cx43) is the most abundant and ubiquitously distributed gap junction protein in testicular cells. Lack of Cx43 expression results in male infertility. We investigated whether Cx43 is expressed and regulated in Leydig, Sertoli and germinal cells at different stages of postnatal development. Cx43 was detected using three different antibodies shown by immunoblotting to be highly specific. At different postnatal ages Cx43 localization was compared in serial or double labeled testicular cryosections with immunocytochemical distribution of steroidogenic enzyme, 3 ␤hidroxysteroid-dehydrogenase (3␤HSD), Mullerian inhibitory hormone (MIH), and germinal nuclear cell antigen (GNCA1), which are specific markers of interstitial Leydig, Sertoli and germinal cells, respectively. In the interstitium, round cell clumps (RCC) with lipid droplets positive for 3␤HSD and Cx43 were frequently found at intertubular areas at birth and Cx43 was mainly localized at cell membrane appositions. From day 3, the number and size of 3␤HSD-positive RCC started to decrease, and reached a minimum at 7-14 dpp; Cx43 expressed by them is progressively downregulated. From day 21 an increase in the size and number of RCC positive for Cx43 and 3␤HSD was found that continued at 24, 26 and 28 days and reached a maximum at 35 and 60 dpp. Biphasic expression of interstitial Cx43 and 3␤HSD was also found to be positively and temporally correlated with fluctuations in intratesticular testosterone content at all ages studied. In the seminiferous cord (SC), Cx43 was expressed at birth between adjacent Sertoli cells (MIH positive) localized at the periphery, as well as in their cytoplasm projections that surround centrally localized gonocytes. From days 3 to 7, Cx43 labeling increased in Sertoli cells mainly at their apical border. At day 14, Cx43 distribution in Sertoli cells changed from apical to basal in parallel to migration of germinal (GNCA1-positive) cells from the periphery to the center of the SC. At all these ages, Cx43 was also localized at cell borders between Sertoli and germinal cells. In conclusion, this study demonstrates that Cx43 in Leydig cells is regulated during postnatal development in an age and functional dependent manner. In the tubule, it is demonstrated that Cx43 is modulated in Sertoli cells during the neonatal and prepubertal period. We also provide evidence for the first time that Cx43-gap junctions communicate between Sertoli and germinal cells before and during the first wave of spermatogenesis. Anat Rec 264: [13][14][15][16][17][18][19][20][21][22][23][24] 2001.

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Developmental regulation of connexin 43 expression in fetal mouse testicular cells

Pérez-Armendáriz

Lamoyi²,

Mason

et al. 2001

Anat. Rec.

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Multiple connexins have been identified in testicular cells. Several lines of evidences indicate that, among them, connexin 43 (Cx43) may be unique for control of gonad development and spermatogenesis. To date, however, it is not known whether Cx43 is expressed in the fetal testis and what possible types of cellular interactions mediated by this connexin are critical to male fertility. In the present work, expression of Cx43 was investigated at various developmental ages in cryosections from mouse testis by using specific antibodies against Cx43. In serial or double-labeled sections, Cx43 localization was compared with immunocytochemical distribution of steroidogenic enzyme, 3␤-hydroxysteroid dehydrogenase (3␤HSD), Mullerian inhibitory hormone (MIH), and germinal nuclear cell antigen (GCNA1), which are specific markers, respectively, of interstitial Leydig, Sertoli, and germinal cells. Sections were analyzed by fluorescence microcopy. We found that Cx43 immunofluorescence (IF) was uniformly distributed in the undifferentiated gonad at 11.5 days post coitus (dpc) and in cells of the mesonephric tubules. In the undifferentiated gonad, Cx43 was localized between primordial germ cells and somatic cells. At 12.5 dpc, when the gonad has undergone sexual differentiation, in the interstitium Cx43 was localized in Leydig cells and in the seminiferous cord it was localized between adjacent Sertoli cells. In Leydig and Sertoli cells, Cx43 labeling increased at 14.5, 16.5, and 18.5 dpc. From day 12.5 up to 18.5 dpc, Cx43 was also localized in cell borders between germinal and Sertoli cells. In conclusion, this study demonstrates that from the earliest stages of gonadal development, Cx43 is expressed in the principal cell types that participate in the control of male fertility. It also shows that Cx43 expression in Leydig and Sertoli cells increase during fetal life. Finally, it provides evidence that, throughout embryonic life, Cx43 forms gap junctions between Sertoli and germinal cells. Anat Rec 264: [237][238][239][240][241][242][243][244][245][246] 2001.

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Characterization of gap junctions between pairs of Leydig cells from mouse testis

Cited by 61 publications

References 24 publications

Peptidergic not monoaminergic fibers profusely innervate the young adult human testis

Peptidergic not monoaminergic fibers profusely innervate the young adult human testis

Expression of Connexin43 in mouse Leydig, Sertoli, and germinal cells at different stages of postnatal development

Developmental regulation of connexin 43 expression in fetal mouse testicular cells

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