[Show abstract][Hide abstract] ABSTRACT: The estrogen 17β-estradiol (E) increases the axospinous synaptic density and plasticity in the hippocampal CA1 region of young female rats but fails to do so in aged female rats. This E stimulus on synaptic plasticity is associated with the phosphorylation-dependent activation of Akt kinase. Our previous findings demonstrated that increased estrogen levels subsequently increase phosphorylated Akt (pAkt)-immunoreactivity (-IR) within the dendritic shafts and spines of pyramidal neurons in young female rats. Therefore, because Akt can promote cell survival and growth, we tested the hypothesis that the less plastic synapses of aged female rats would contain less E-stimulated pAkt-IR. Here, young (3-4 months) and aged (22-23 months) female rats were ovariectomized 7 days prior to a 48-h administration of either vehicle or E. The pAkt-IR synaptic distribution was then analyzed using post-embedding electron microscopy. In both young and aged rats, pAkt-IR was found in dendritic spines and terminals, and pAkt-IR was particularly abundant at the post-synaptic density. Quantitative analyses revealed that the percentage of pAkt-labeled synapses was significantly greater in young rats compared to aged rats. Nonetheless, E treatment significantly increased pAkt-IR in pre- and post-synaptic profiles of both young and aged rats, although the stimulus in young rats was notably more widespread. These data support the evidence that hormone-activated signaling associated with cell growth and survival is diminished in the aged brain. However, the observation that E can still increase pAkt-IR in aged synapses presents this signaling component as a candidate target for hormone replacement therapies.
Brain research 03/2011; 1379:98-108. DOI:10.1016/j.brainres.2010.07.053 · 2.84 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Physical activity has a modulatory role on regulatory steps of excitation-contraction coupling (ECC) determining skeletal muscle contractility. We evaluated and compared the contractile responsiveness and caffeine-induced contractures of fast (extensor digitorum longus; EDL) and slow-twitch (soleus; SOL) muscles in suspension hypokinesia (SH) and exercised rats. After SH or low intensity exercise, EDL and SOL were isolated, twitch and tetanic contractions and caffeine (10 mM) contractures were recorded. Twitch and tetanic contractions of EDL increased by 60% in exercised rats (p <0.05) while no alteration was observed after SH. Exercise did not alter twitch and tetanic contractions of SOL, while SH depressed contractions (p <0.05). Caffeine contractures were diminished in exercised rat EDL (P <0.05). In SH-rat EDL, contractures increased in amplitude (p <0.01) with a rapid time course (p <0.05). Contractures did not change in SOL after exercise or SH. We concluded that SH and exercise exerted diverse modulatory effects on skeletal muscle contractility. Contractile improvement due to exercise was prominent in EDL. Our results suggest that the muscle-type specific adaptations are related to a change in ECC due to the differences in the regulatory steps, particularly in the intracellular Ca(2+) handling mechanisms.
[Show abstract][Hide abstract] ABSTRACT: Estradiol (E) mediates increased synaptogenesis in the hippocampal CA1 stratum radiatum (sr) and enhances memory in young and some aged female rats, depending on dose and age. Young female rats express more estrogen receptor α (ERα) immunolabeling in CA1sr spine synapse complexes than aged rats and ERα regulation is E sensitive in young but not aged rats. The current study examined whether estrogen receptor β (ERβ) expression in spine synapse complexes may be altered by age or E treatment. Young (3-4 months) and aged (22-23 months) female rats were ovariectomized 7 days prior to implantation of silastic capsules containing either vehicle (cholesterol) or E (10% in cholesterol) for 2 days. ERβ immunoreactivity (ir) in CA1sr was quantitatively analyzed using post-embedding electron microscopy. ERβ-ir was more prominent post-synaptically than pre-synaptically and both age and E treatment affected its synaptic distribution. While age decreased the spine synaptic complex localization of ERβ-ir (i.e., within 60 nm of the pre- and post-synaptic membranes), E treatment increased synaptic ERβ in both young and aged rats. In addition, the E treatment, but not age, increased dendritic shaft labeling. This data demonstrates that like ERα the levels of ERβ-ir decrease in CA1 axospinous synapses with age, however, unlike ERα the levels of ERβ-ir increase in these synapses in both young and aged rats in response to E. This suggests that synaptic ERβ may be a more responsive target to E, particularly in aged females.
Brain research 09/2010; 1379:86-97. DOI:10.1016/j.brainres.2010.09.069 · 2.84 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Clustered protocadherins (Pcdhs) are a family of cadherin-like molecules arranged in gene clusters (α, β, and γ). γ-Protocadherins
(Pcdh-γs) are involved in cell-cell interactions, but their prominent intracellular distribution in vivo and different knock-out phenotypes suggest that these molecules participate in still unidentified processes. We found using
correlative light and electron microscopy that Pcdh-γA3 and -γB2, but not -γC4, -α1, or N-cadherin, generate intracellular
juxtanuclear membrane tubules when expressed in cells. These tubules recruit the autophagy marker MAP1A/1B LC3 (LC3) but are
not associated with autophagic vesicles. Lipidation of LC3 is required for its coclustering with Pcdh-γ tubules, suggesting
the involvement of an autophagic-like molecular cascade. Expression of wild-type LC3 with Pcdh-γA3 increased tubule length
whereas expression of lipidation-defective LC3 decreased tubule length relative to Pcdh-γA3 expressed alone. The tubules were
found to emanate from lysosomes. Deletion of the luminal/extracellular domain of Pcdh-γA3 preserved lysosomal targeting but
eliminated tubule formation whereas cytoplasmic deletion eliminated both lysosomal targeting and tubule formation. Deletion
of the membrane-proximal three cadherin repeats resulted in tubes that were narrower than those produced by full-length molecules.
These results suggest that Pcdh-γA and -γB families can influence the shape of intracellular membranes by mediating intraluminal
interactions within organelles.
Journal of Biological Chemistry 07/2010; 285(27):20982-20992. · 4.57 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Clustered protocadherins (Pcdhs) are a family of cadherin-like molecules arranged in gene clusters (alpha, beta, and gamma). gamma-Protocadherins (Pcdh-gammas) are involved in cell-cell interactions, but their prominent intracellular distribution in vivo and different knock-out phenotypes suggest that these molecules participate in still unidentified processes. We found using correlative light and electron microscopy that Pcdh-gammaA3 and -gammaB2, but not -gammaC4, -alpha1, or N-cadherin, generate intracellular juxtanuclear membrane tubules when expressed in cells. These tubules recruit the autophagy marker MAP1A/1B LC3 (LC3) but are not associated with autophagic vesicles. Lipidation of LC3 is required for its coclustering with Pcdh-gamma tubules, suggesting the involvement of an autophagic-like molecular cascade. Expression of wild-type LC3 with Pcdh-gammaA3 increased tubule length whereas expression of lipidation-defective LC3 decreased tubule length relative to Pcdh-gammaA3 expressed alone. The tubules were found to emanate from lysosomes. Deletion of the luminal/extracellular domain of Pcdh-gammaA3 preserved lysosomal targeting but eliminated tubule formation whereas cytoplasmic deletion eliminated both lysosomal targeting and tubule formation. Deletion of the membrane-proximal three cadherin repeats resulted in tubes that were narrower than those produced by full-length molecules. These results suggest that Pcdh-gammaA and -gammaB families can influence the shape of intracellular membranes by mediating intraluminal interactions within organelles.
[Show abstract][Hide abstract] ABSTRACT: Hippocampal dendritic spine and synapse numbers in female rats vary across the estrous cycle and following experimental manipulation of hormone levels in adulthood. Based on behavioral studies demonstrating that learning patterns are altered following puberty, we hypothesized that dendritic spine number in rat hippocampal CA1 region would change postpubertally. Female Sprague-Dawley rats were divided into prepubertal (postnatal day (P) 22), peripubertal (P35) and postpubertal (P49) groups, with the progression of puberty evaluated by vaginal opening, and estrous cyclicity subsequently assessed by daily vaginal smears. Spinophilin immunoreactivity in dendritic spines was used as an index of spinogenesis in area CA1 stratum radiatum (CA1sr) of hippocampus. First, electron microscopy analyses confirmed the presence of spinophilin specifically in dendritic spines of CA1sr, supporting spinophilin as a reliable marker of hippocampal spines in young female rats. Second, stereologic analysis was performed to assess the total number of spinophilin-immunoreactive puncta (i.e. spines) and CA1sr volume in developing rats. Our results indicated that the number of spinophilin-immunoreactive spines in CA1sr was decreased 46% in the postpubertal group compared to the two younger groups, whereas the volume of the hippocampus underwent an overall increase during this same developmental time frame. Third, to determine a potential role of estradiol in this process, an additional group of rats was ovariectomized (OVX) prepubertally at P22, then treated with estradiol or vehicle at P35, and spinophilin quantified as above in rats perfused on P49. No difference in spinophilin puncta number was found in OVX rats between the two hormone groups, suggesting that this developmental decrease is independent of peripheral estradiol. These changes in spine density coincident with puberty may be related to altered hippocampal plasticity and synaptic consolidation at this phase of maturity.
[Show abstract][Hide abstract] ABSTRACT: 17beta-Estradiol (E) increases axospinous synapse density in the hippocampal CA1 region of young female rats, but not in aged rats. This may be linked to age-related alterations in signaling pathways activated by synaptic estrogen receptor alpha (ER-alpha) that potentially regulate spine formation, such as LIM-kinase (LIMK), an actin depolymerizing factor/cofilin kinase. We hypothesized that, as with ER-alpha, phospho-LIM-kinase (pLIMK) may be less abundant or responsive to E in CA1 synapses of aged female rats. To address this, cellular and subcellular distribution of pLIMK-immunoreactivity (IR) in CA1 was analyzed by light and electron microscopy in young and aged female rats that were ovariectomized and treated with either vehicle or E. pLIMK-IR was found primarily in perikarya within the pyramidal cell layer and dendritic shafts and spines in stratum radiatum (SR). While pLIMK-IR was occasionally present in terminals, post-embedding quantitative analysis of SR showed that pLIMK had a predominant post-synaptic localization and was preferentially localized within the postsynaptic density (PSD). The percentage of pLIMK-labeled synapses increased (30%) with E treatment (P<0.02) in young animals, and decreased (43%) with age (P<0.002) regardless of treatment. The pattern of distribution of pLIMK-IR within dendritic spines and synapses was unaffected by age or E treatment, with the exception of an E-induced increase in the non-synaptic core of spines in young females. These data suggest that age-related synaptic alterations similar to those seen with ER-alpha occur with signaling molecules such as pLIMK, and support the hypothesis that age-related failure of E treatment to increase synapse number in CA1 may be due to changes in the molecular profile of axospinous synapses with respect to signaling pathways linked to formation of additional spines and synapses in response to E.
[Show abstract][Hide abstract] ABSTRACT: Responses of cremaster muscles and sacs from boys with undescended testis suggested less exposure against sympathetic, but more exposure against parasympathetic tonuses. Since the sympathetic tonus is androgen dependent, it has been suggested that androgens control the descent by influencing the sympathetic tonus. Therefore an experimental study was planned to evaluate the contractile responses of cremaster muscles according to locations of associated testes in rats subjected to intrauterine steroidal or non-steroidal anti-androgen exposure.
Time-mated pregnancies were started in 18 rats. They were divided into three groups and each group was given physiologic saline, flutamide or cyproterone acetate (Androcur Depot). Flutamide was administered from day 15 to day 19, cyproterone acetate on day 15 of intra-uterine life. At twelve weeks of age the localization of testes was evaluated, cremaster muscles were removed, and contractile properties were studied. Twitch and tetanic contractions were recorded isometrically at 37 degrees C. Effects of verapamil and isoprenaline were investigated. Results were compared by ANOVA and p values less than 0.05 were considered significant.
Both testes of all available male offspring in the saline (n = 22) and cyproterone acetate-treated (n = 19) groups were in the scrotum. Sixty percent of males in the flutamide-treated group (n = 20) had undescended testes. Cremaster muscles of rats exposed to flutamide had a lower sensitivity to voltage sensitive Ca+2 channel blockade by verapamil (3 x 10(-4) mol/L, p < 0.05) and displayed greater contractile response to isoprenaline (10(-5) mol/L, p < 0.05). Alterations in contractile properties of the muscles did not differ according to localization of testes in rats subjected to flutamide exposure.
CM in rats subjected to non-steroidal anti-androgen exposure revealed alterations that indicate a decrease in sympathetic tonus. Since non-steroidal anti-androgen also inhibits the descent, the present study provides experimental support for the involvement of sympathetic tonus in the androgenic control of testicular descent.
European Journal of Pediatric Surgery 09/2005; 15(4):273-8. DOI:10.1055/s-2005-837625 · 0.99 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Androgens are proposed to influence testicular descent through modulating sympathetic tone. An experimental study was undertaken to evaluate the effects of prenatal chemical sympathectomy on testicular location associated with the alterations in contractile properties of cremaster muscles in rats.
Time-mated pregnancies were started in 10 rats. Two groups, each receiving saline or 6-hydroxydopamine from day 15 to day 19 of intrauterine life were established. At 2 months of age, localization of testes were evaluated, cremaster muscles were removed, and contractile properties were studied. Twitch and tetanic contractions were recorded isometrically at 37 degrees C. Effects of verapamil, isoprenaline, and L-NNA were investigated. Results were compared through analysis of variance (ANOVA), and P values less than.05 were considered to be significant.
Both testes of all male offspring in the control group (n = 19) were in the scrotum. Six offspring among 17 subjected to 6-hydroxydopamine had undescended testes. Treatment with 6-hydroxydopamine had no effect on force-frequency relationship of cremaster muscle strips. Cremaster muscles of rats exposed to 6-hydroxydopamine had lower sensitivity to voltage-sensitive Ca++ channel blockade by verapamil (3 x 10(4) mol/L; P <.05). These muscles displayed greater contractile response to isoprenaline (10(-5) mol/L; P <.05) but not to nitric oxide synthase inhibition by N(omega)-nitro-L-arginine. Alterations in contractile properties of the muscles did not differ according to localization of testes among rats subjected to 6-hydroxydopamine.
Administration of 6-hydroxydopamine resulted in suprascrotally located testes. This localization has been associated with less exposure at sympathetic tonus. These findings support that sympathetic activity plays an important role in localization of testis.
Journal of Pediatric Surgery 11/2003; 38(11):1628-32. DOI:10.1016/S0022-3468(03)00574-8 · 1.39 Impact Factor