The gut microbiome restores intrinsic and extrinsic nerve function in germ-free mice accompanied by changes in calbindin.
BACKGROUND
The microbiome is essential for normal myenteric intrinsic primary afferent neuron (IPAN) excitability. These neurons control gut motility and modulate gut-brain signaling by exciting extrinsic afferent fibers innervating the enteric nervous system via an IPAN to extrinsic fiber sensory synapse. We investigated effects of germ-free (GF) status and conventionalization on extrinsic sensory fiber discharge in the mesenteric nerve bundle and IPAN electrophysiology, and compared these findings with those from specific pathogen-free (SPF) mice. As we have previously shown that the IPAN calcium-dependent slow afterhyperpolarization (sAHP) is enhanced in GF mice, we also examined the expression of the calcium-binding protein calbindin in these neurons in these different animal groups.


METHODS
IPAN sAHP and mesenteric nerve multiunit discharge were recorded using ex vivo jejunal gut segments from SPF, GF, or conventionalized (CONV) mice. IPANs were excited by adding 5 μM TRAM-34 to the serosal superfusate. We probed for calbindin expression using immunohistochemical techniques.


RESULTS
SPF mice had a 21% increase in mesenteric nerve multiunit firing rate and CONV mice a 41% increase when IPANs were excited by TRAM-34. For GF mice, this increase was barely detectable (2%). TRAM-34 changed sAHP area under the curve by -77 for SPF, +3 for GF, or -54% for CONV animals. Calbindin-immunopositive neurons per myenteric ganglion were 36% in SPF, 24% in GF, and 52% in CONV animals.


CONCLUSIONS
The intact microbiome is essential for normal intrinsic and extrinsic nerve function and gut-brain signaling.
McMaster Brain-Body Institute at St Joseph's Healthcare, Hamilton, ON, Canada; Department of Psychiatry and Behavioral Neurosciences, McMaster University, Hamilton, ON, Canada.
K A
KA
McVey Neufeld
A
A
Perez-Burgos
Y K
YK
Mao
J
J
Bienenstock
W A
WA
Kunze
D000200:Action Potentials / Q000187:drug effects / Q000502:physiology*; D000818:Animals; D064026:Calbindins / Q000378:metabolism*; D017615:Enteric Nervous System / Q000187:drug effects / Q000378:metabolism / Q000502:physiology; D005260:Female; D000069196:Gastrointestinal Microbiome / Q000502:physiology*; D005856:Germ-Free Life; D007583:Jejunum / Q000294:innervation* / Q000378:metabolism / Q000382:microbiology; D051379:Mice; D009197:Myenteric Plexus / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D009475:Neurons, Afferent / Q000187:drug effects / Q000502:physiology*; D011720:Pyrazoles / Q000494:pharmacology; D013047:Specific Pathogen-Free Organisms
D064026:Calbindins; D011720:Pyrazoles; C411671:TRAM 34
TRAM-34; calbindin; enteric nervous system; germ-free; mesenteric afferent nerve; microbiota