Modulation of GAP-43 mRNA by GABA and glutamate in cultured cerebellar granule cells.

Expression of GAP-43 in the cerebellum and selected regions of the brain has been shown to be developmentally regulated. Localization of GAP-43 mRNA within granule cells of the immature and mature rat cerebellum has been demonstrated by in situ hybridization. Higher levels are detected in the neonate compared to the adult.

Distribution of GAP-43 mRNA in the immature and adult cerebellum: a role for GAP-43 in cerebellar development and neuroplasticity.

Expression of GAP-43 mRNA in the rat cerebellum and inferior olivary nucleus was examined at birth, during postnatal development and in the adult by both Northern and in situ hybridization. Northern blot analysis revealed that cerebellar GAP-43 mRNA expression increases from birth to postnatal day (PD) 7 and then declines to a lower level in the adult. At birth, in situ hybridization experiments showed intense labeling of GAP-43 mRNA in the premigratory, but not the germinal, zone of the cerebellar external granule cell layer.

Particle bombardment: a universal approach for gene transfer to cells and tissues.

In the past year, significant progress in the field of gene transfer has been made possible by refinement of the technique of particle bombardment. The process has been utilized for the study of gene expression in plastids and mitochondria, the production of transgenic crop plants and gene transfer into live animals. Bombarding tissues of live animals with genes that code for antigenic proteins may provide an effective means of vaccination.

Gene transfer to tumor-infiltrating lymphocytes and other mammalian somatic cells by microprojectile bombardment.

We demonstrate the application of particle delivery to the transformation of mammalian somatic cells. Two mouse T-lymphocyte cell lines and Chinese hamster ovary (CHO) cells were transformed transiently with the RSV-ADH (alcohol dehydrogenase) gene. Stable gene transfer was also demonstrated in CHO cells at a frequency of 6 x 10(-4) and in T-cells at a frequency of 1 x 10(-4), using beta-Gal/neomycin-resistance gene constructs.

Transformation of microbes, plants and animals by particle bombardment.

Over the past several years, particle bombardment has evolved into a useful tool for molecular biologists, allowing direct gene transfer to a broad range of cell and tissue types. Some of the important applications of the process include the production of transgenic crop species including maize and soybean and the introduction of DNA into plastids and mitochondria. Recent results have extended the range of gene transfer by particle bombardment to animal and bacterial cells.

Ontogeny of somatostatin mRNA-containing perikarya in the rat central nervous system.

The distribution of neuronal perikarya containing somatostatin mRNA in the developing rat brain was investigated with in situ hybridization histochemistry. This study describes the expression of somatostatin mRNA during selected perinatal stages and demonstrates regional changes in somatostatin mRNA expression at the single cell level. The mRNA expression closely parallels previously reported developmental localization of the peptide (Inagaki et al.

Light microscopic localization of glycosyltransferase activities in cells and tissues.

We describe an assay for light microscopic visualization of specific glycosyltransferases on tissue sections or on cells. The assay uses a sequence of enzyme reactions that yields two moles of NADH for each mole of the uridine-5'-diphosphate (UDP) released during transfer of a monosaccharide from a UDP sugar to an acceptor. When diaphorase and tetrazolium salts are present in the incubation mixture, the tetrazolium salts are reduced to colored diformazans, which precipitate at the sites of glycosyltransferase activity.

Localization of VIP and PHI-27 messenger RNA in rat thalamic and cortical neurons.

Messenger RNA (mRNA) coding for vasoactive intestinal polypeptide (VIP) and peptide histidine isoleucine (PHI-27) were localized in cortical and thalamic neurons with synthetic DNA probes complementary to the PHI-27 and VIP exon coding sequence of the rat VIP precursor gene. Hybridization signal with these probes was found widely distributed in the thalamus, neocortex, and pyriform cortex, and the distribution of hybridization signal for each probe was identical. Furthermore, the distribution of each message was correlated closely with peptide distribution demonstrated immunohistochemically.

Coordinate hormonal and synaptic regulation of vasopressin messenger RNA.

Previous studies have shown that adrenalectomy augments arginine vasopressin (AVP) messenger RNA levels in the adult paraventricular nucleus. It is now demonstrated that unilateral lesions in the lateral septal nucleus enhance the adrenalectomy-induced expression of AVP mRNA. This effect was entirely ipsilateral to the lesion and most prominent in the rostral paraventricular nucleus and related nuclei.

Neuronal localization of prosomatostatin mRNA in the rat brain with in situ hybridization histochemistry.

Individual neurons containing prosomatostatin mRNA were identified with in situ hybridization histochemistry. Our results demonstrate a widespread distribution of prosomatostatin mRNA in several regions of the rat central nervous system. Neurons containing this transcript were most abundant in the anterior olfactory nucleus, hypothalamus, hippocampus, and amygdala as well as in all regions of the cerebral cortex.

Localization of vasopressin-, vasoactive intestinal polypeptide-, peptide histidine isoleucine- and somatostatin-mRNA in rat suprachiasmatic nucleus.

Messenger RNAs (mRNA) coding for vasoactive intestinal polypeptide (VIP), peptide histidine isoleucine (PHI), somatostatin and vasopressin were localized in the suprachiasmatic nucleus (SCN) of the rat hypothalamus using in situ hybridization histochemistry. Specific mRNA coding for each of these peptides was distributed in areas coextensive with the immunohistochemical localization of the appropriate peptide. The autoradiographic signal produced with probes to VIP and PHI created dense concentrations of silver grains over neuronal perikarya in the ventrolateral SCN, and the coextensive distribution of both VIP- and PHI-mRNAs suggests that both peptides are synthesized within the same neurons.

Hormonal regulation of somatostatin messenger RNA.

The ability of gonadal steroids to regulate the expression of the somatostatin gene in several regions of the CNS was investigated with in situ hybridization histochemistry. The amount of somatostatin mRNA was found to be significantly decreased 2-3 weeks after ovariectomy or orchidectomy in the periventricular hypothalamus, the ventromedial nucleus of the hypothalamus, and the medial and central nuclei of the amygdala. Treatment of gonadectomized rats with estradiol benzoate or testosterone enanthate reversed this decrease in somatostatin mRNA.

Appearance and localization of acetylcholinesterase in embryos of the leech Helobdella triserialis.

The appearance and localization of acetylcholinesterase (AchE) were studied during embryonic development of the leech Helobdella triserialis. Use of a histochemical stain showed that AchE is present in the polar plasms of the uncleaved leech egg. in the course of the first two cleavages, the AchE is transmitted along with the polar plasms mainly to the D blastomere, precursor of mesoderm and ectoderm.