No abstract

Insulin has been identified in the central nervous system of a number of vertebrate species, but the site of synthesis as yet remains unresolved. Two previous studies reported the presence of insulin mRNA in neural tissue, but related efforts to confirm and better localize the cellular origin of the hormone have yielded equivocal results. In the present study we have attempted to clarify this issue by employing both immunocytochemistry and in situ hybridization on isolated enriched cultures of rabbit brain neurons and glia. Our data show that a subset (3-5%) of neurons is positively immunoreactive for insulin, but all of the glial cells are negative. The level of staining intensity can be increased by preincubating the neurons with monensin (a Na+ ionophore that prevents cell secretory activity), but not the fraction of positive cells. Similarly, in situ hybridization reveals the presence of mRNA in 3-5% of neurons, but no such signal is detected in glia. Thus, our data not only confirm previous reports of insulin in the central nervous system, but, more importantly, indicate that the synthesis of the hormone is local and apparently confined to a subset of neurons.

We investigated the presence of three specific types of glucose transporters (GT) within the rabbit central nervous system during various developmental stages. Employing the Hep G2/brain-type insulin-insensitive and the insulin-responsive (IRGT; adipocyte/skeletal muscle type) GT antibody and cDNA, we studied protein and mRNA within the whole brain (25-, 27-, and 30-day-old fetus; 1-, 5-, 10-day-old neonate; and adult), using cultured neuronal and glial cells, by Western and Northern blot analysis. Similarly, using the insulin-insensitive human fetal skeletal muscle-type (GLUT-3) GT cDNA, we characterized this mRNA by Northern blot analysis. Additional confirmation of cell specificity was sought by performing immunohistochemical staining on the neuronal and glial cells to detect the specific type of GT protein. We observed a developmental regulation of brain-type GT within the whole brain, the peak abundance of protein and mRNA occurring in the adult, followed next by the fetus. No IRGT was detected within the whole brain at any stage of development. Contrary to the brain-type GT mRNA, GLUT-3 mRNA was found to be most abundant in the 10-day-old neonate and adult, followed next by the early neonate, with little in the fetus. Within isolated brain cell cultures, the mRNAs for the brain- and GLUT-3-types of GTs were abundantly present within glial cells, with considerably lesser amounts noted within the neurons. IRGT, on the other hand, revealed rather weak mRNA bands in both glial and neuronal cells. Western blotting revealed a brain type of GT protein within the glial cells alone; the neuronal cells for the most part were devoid of both the brain-type and the IRGT proteins. Further immunohistochemical staining confirmed the definite presence of the brain-type GT within the glial cells, with slight immunoreactivity observed within the neurons. Additionally, no significant IRGT immunoreactivity was observed within either cell type. We did not study the GLUT-3 type of immunoreactivity within neurons and glia. We conclude that both the Hep G2/brain and the GLUT-3 types, and not the IRGT, are developmentally regulated within the whole brain. Further, the Hep G2/brain and the GLUT-3 types of GTs are distinctly present within glial cells, with none to minute amounts present within the neurons. No IRGT protein is observed within the whole brain and the two cell types. These results suggest a differential expression of specific GT types within the neuronal and glial components of the brain.

Background The addition of budesonide to surfactant in very-low-birth-weight infants with less severe RDS decreased bronchopulmonary dysplasia (BPD) severity. Long-term neurodevelopmental follow-up was needed to monitor for systemic effects of budesonide. Methods Infants ≤1250 g who received intratracheal budesonide (0.25 mg/kg) with surfactant ( n = 173) were compared to a historical cohort who received surfactant alone ( n = 294). Peabody Developmental Motor Scales II at 4–6 months corrected age and Bayley Scales of Infant & Toddler Development III at 18–22 months corrected age were compared. Results There were no differences in muscle tone or motor skills by Peabody exam. There were no differences in the cognitive, language, or motor domains between cohorts on Bayley III. Conclusions In a cohort of infants treated with budesonide mixed with surfactant, there were no differences in developmental outcomes at 4–6 months or 18–22 months corrected age.