Search results for "Labeling"
showing 10 items of 372 documents
Toward an integrative and predictive sperm quality analysis in Bos taurus
2017
There is a need to develop more integrative sperm quality analysis methods, enabling researchers to evaluate different parameters simultaneously cell by cell. In this work, we present a new multi-parametric fluorescent test able to discriminate different sperm subpopulations based on their labeling pattern and motility characteristics. Cryopreserved semen samples from 20 Holstein bulls were used in the study. Analyses of sperm motility using computer-assisted sperm analysis (CASA-mot), membrane integrity by acridine orange-propidium iodide combination and multi-parametric by the ISAS®3Fun kit, were performed. The new method allows a clear discrimination of sperm subpopulations based on memb…
Neurochemistry of olivocochlear neurons in the hamster.
2009
The present study was conducted to characterize the superior olivary complex (SOC) of the lower brain stem in the pigmented Djungarian hamster Phodopus sungorus. Using Nissl-stained serial cryostat sections from fresh-frozen brains, we determined the borders of the SOC nuclei. We also identified olivocochlear (OC) neurons by retrograde neuronal tracing upon injection of Fluoro-Gold into the scala tympani. To evaluate the SOC as a putative source of neuronal nitric oxide synthase (nNOS), arginine-vasopressin (AVP), oxytocin (OT), vasoactive intestinal polypeptide (VIP), or pituitary adenylate cyclase-activating polypeptide (PACAP) that were all found in the cochlea, we conducted immunohistoc…
A nitrergic projection from the superior olivary complex to the inferior colliculus of the rat
2003
The present study was conducted to test whether the ascending auditory projection from the superior olivary complex (SOC) of the brainstem to the inferior colliculus (IC) may use nitric oxide (NO) as a neuroactive compound. We identified olivo-collicular projection neurons in subnuclei of the SOC by retrograde neuronal tracing with Fluoro-Gold (FG) injected into the central nucleus of the IC. Sections containing retrograde labelled neurons were subjected to immunohistochemical incubation in an antiserum directed against the enzyme responsible for NO production in nerve cells, neuronal NO synthase (nNOS). The analysis showed that FG-containing neurons as well as nNOS-immunoreactive neurons w…
Infrequent co-existence of nitric oxide synthase and parvalbumin, calbindin and calretinin immunoreactivity in rat pontine neurons.
1995
Neurons in the laterodorsal tegmental nucleus (LDTg), ventrolateral dorsal tegmental nucleus (LDTgV), pedunculopontine tegmental nucleus (PPTg), lateral and medial parabrachial nuclei (LPB and MPB) were immunoreactive to brain nitric oxide synthase (NOS) or isoform I. Double-labeling experiments showed that very few NOS-containing neurons in the pons were immunoreactive to any of the three calcium-binding proteins: calbindin-D 28K (CB-IR), parvalbumin (PV-IR) and calretinin (CR-IR). These findings extend our previous observation in the neocortex and suggest that a population of central NOS-containing neurons can be neurochemically characterized as CB/CR/PV deficient.
Botulinum Toxin Type B Blocks Sudomotor Function Effectively: A 6 Month Follow Up
2003
This study analyzes the suppression of sweat gland activity by botulinum toxin type B. We injected botulinum toxin type B (between 2 and 1000 mouse units subcutaneously) in the lateral side of both lower legs in 15 healthy volunteers. Sweat tests were carried out before botulinum toxin type B injections, and at 3 wk, 3 mo, and 6 mo. We studied focal anhidrosis by iodine–starch staining and by capacitance hygrometry after carbachol iontophoresis, according to the quantitative sudomotor axon reflex test (QSART). Iodine starch staining indicated that a threshold dose of 8 mouse units botulinum toxin type B leads to anhidrotic skin spots (>4 cm2) after 3 wk. Duration of anhidrosis was prolonged…
Pregnenolone sulfate, a naturally occurring excitotoxin involved in delayed retinal cell death.
2002
The present study was designed to investigate the neurosteroid pregnenolone sulfate (PS), known for its ability to modulate NMDA receptors and interfere with acute excitotoxicity, in delayed retinal cell death. Three hours after exposure of the isolated and intact retina to a 30-min PS pulse, DNA fragmentation as assessed by genomic DNA gel electrophoresis and a modified in situ terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling (TUNEL) method appeared concurrently with an increase in superoxide dismutase (SOD) activity and thiobarbituric acid-reactive substances (TBARS) levels. At 7 h, the increased amount of DNA laddering was accompanied by a higher number of TUN…
Spermatocyte chromosome analysis of Helicella virgata (Pulmonata: Helicidae): silver-stained and C-banded chromosomes.
1991
Chromosome numbers of the snail Helicella virgata from the fields of Castellammare del Golfo (Sicily) are n = 26 and 2n = 52. Silver-staining analyses of testicular cells suggest that both mitotic and meiotic chromosomes are involved in nucleolus organization. A within-individual variability in NOR-banding pattern is present in each of the 20 specimens analyzed.
Identification of calcium sensing receptor (CaSR) mRNA-expressing cells in normal and injured rat brain
2009
Calcium sensing receptor (CaSR), isolated for the first time from bovine and human parathyroid, is a G-protein-coupled receptors that has been involved in diverse physiological functions. At present a complete in vivo work on the identification of CaSR mRNA-expressing cells in the adult brain lacks and this investigation was undertaken in order to acquire more information on cell type expressing CaSR mRNA in the rat brain and to analyse for the first time its expression in different experimental models of brain injury. The expression of CaSR mRNAs was found mainly in scattered cells throughout almost all the brain regions. A double labeling analysis showed a colocalization of CaSR mRNA expr…
In vitro modeling of the ternary interaction in juvenile hormone metabolism
1996
The gradual decline in juvenile hormone (JH) titer followed by its complete clearance early in the last larval instar is required for the onset of the metamorphosis of lepidopterous larvae. JH titer is regulated by both biosynthesis and degradation. Two major pathways for JH metabolism, ester hydrolysis and epoxide hydration, are due to JH esterase (JHE) and JH epoxide hydrolase (JHEH), respectively. In vitro experiments designed to elucidate the molecular mechanism of JH metabolism are described. First, microsomal JHEH in Manduca sexta eggs was identified by using photoaffinity analogs of JH, and purified to homogeneity with ion exchange and hydroxylapatite columns. Purified JHEH from M. s…
Purification and reassessment of ligand binding by the recombinant, putative juvenile hormone receptor of the tobacco hornworm, Manduca sexta
1996
The 29 kDa protein from the larval epidermis of the tobacco hornworm, Manduca sexta, that specifically bound photoaffinity analogs of JH I and JH II was produced by a recombinant baculovirus (rJP29). The higher of the two molecular weight forms made corresponded to a protein that could be formed by read‐through of the TGA termination codon to the following TAA. The previously reported, apparent high affinity binding of [methyl‐3H]‐JH I by rJP29 as measured by the dextran‐coated charcoal (DCC) assay [Palli et al., Proc Natl Acad Sci USA 91:6191–6195 (1994)] was found to be artifactual due to endogenous cellular esterases that co‐purified with rJP29 through both DEAE cellulose and MonoQ chrom…