Age-dependent changes in cholinergic neurons of the rat small and large intestines

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Abstract

BACKGROUND: Neurons containing the acetylcholine-synthesizing enzyme, choline acetyltransferase (ChAT), constitute a significant proportion of cells in the myenteric (MP) and submucosal (SP) plexuses of the small and large intestines. However, current studies lack data on age-related changes in this group of neurons during postnatal ontogenesis.

AIM: To determine postnatal changes in ChAT-containing neurons in the intramural ganglia of the rat intestines.

METHODS: The study was conducted on rats of the following age groups: newborns; animals aged 10, 20, 30, and 60 days; and animals aged 12 and 24 months. Changes in the percentage and average cross-sectional area of ChAT-immunoreactive (IR) neurons in the MP and SP of the small and large intestines of rats during postnatal ontogenesis were analyzed using double-labeling immunohistochemistry with antibodies to ChAT and PGP9.5.

RESULTS: ChAT was detected in the majority of neurons in the MP and SP of the small and large intestines across all age groups. In the MP of the small intestine, the percentage of ChAT-IR neurons increased from the neonatal period to day 20 of life and then gradually decreased until old age. In the MP of the large intestine, the proportion of ChAT-IR neurons increased from the neonatal period to day 10 of life and then also decreased starting from day 20 until old age. In the SP of the small intestine, the percentage of ChAT-IR neurons was higher in newborns compared with 10-day-old rats, and in newborns and day 20 rats compared with aged animals. In the SP of the large intestine, the percentage of ChAT-IR neurons increased during the first 10 days of life, decreased between days 10 and 20, and then remained unchanged at older ages.

No differences were found in the average cross-sectional areas of ChAT-IR and ChAT-negative neurons in the small and large intestines. The average cross-sectional area of ChAT-IR neurons increased from birth to day 60 in both the MP and SP of the small and large intestines.

CONCLUSION: These data indicate age-related changes in the neurotransmitter composition of the intestinal intramural ganglia. During postnatal ontogenesis, the percentage of ChAT-IR neurons in the nerve plexuses of the small and large intestines changes: it increases in the early postnatal period and decreases in old age.

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About the authors

Antonina F. Budnik

Kabardino-Balkarian State University

Email: budnik74@mail.ru
ORCID iD: 0000-0002-3333-5865
SPIN-code: 3691-4817

MD, Cand. Sci. (Medicine), Assistant Professor

Russian Federation, Nalchik

Petr M. Masliukov

Yaroslavl State Medical University

Author for correspondence.
Email: mpm@ysmu.ru
ORCID iD: 0000-0002-6230-5024
SPIN-code: 7676-0849

MD, Dr. Sci. (Medicine), Professor

Russian Federation, Yaroslavl

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2. Fig. 1. Immunohistochemical staining of neurons in the ganglia of the intermuscular plexus of the small intestine in rats: a — 10 days old; b — 20 days old; c — 24 months old; red channel (Cy3) — choline acetyltransferase (CAT), green channel (FITC) — PGP9.5 protein; arrows indicate CAT-negative neurons; scale bar 50 µm.

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3. Fig. 2. Immunohistochemical staining of neurons in the ganglia of the submucosal plexus of the small intestine in rats: a — 10 days old; b — 20 days old; c — 24 months old; red channel (Cy3) — choline acetyltransferase (CAT), green channel (FITC) — PGP9.5 protein; arrows indicate CAT-negative neurons; scale bar 50 µm.

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4. Fig. 3. Immunohistochemical staining of neurons in the ganglia of the intermuscular plexus of the rat colon: a — 10 days old; b — 20 days old; c — 24 months old; red channel (Cy3) — choline acetyltransferase (CAT), green channel (FITC) — PGP9.5 protein; arrows indicate CAT-negative neurons; scale bar 50 µm.

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5. Fig. 4. Immunohistochemical staining of neurons in the ganglia of the submucosal plexus of the rat colon: a — 10 days old; b — 20 days old; c — 24 months old; red channel (Cy3) — choline acetyltransferase (CAT), green channel (FITC) — PGP9.5 protein; arrows indicate CAT-negative neurons; scale bar 50 µm.

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6. Fig. 5. Diagram showing changes in the proportion of neurons containing choline acetyltransferase (CAT) in the ganglia of the intermuscular (IM) and submucosal (SM) plexuses of the small and large intestines of rats during postnatal ontogenesis: * p < 0.05 compared with 20-day-old rats; # p < 0.05 compared with the large intestine.

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7. Fig. 6. Diagram showing changes in the mean cross-sectional area of neurons containing and not containing choline acetyltransferase (CHAT+ and CHAT−, respectively) in the ganglia of the intermuscular (IM) and submucosal (SM) plexuses of the rat intestine during postnatal ontogenesis: a — small intestine; b — large intestine; * p < 0.05 compared with the myenteric plexus.

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