Brett A. Johnson

Regulation of neonatal and adult mammalian heart regeneration by the miR-15 family

We recently identified a brief time period during postnatal development when the mammalian heart retains significant regenerative potential after amputation of the ventricular apex. However, one major unresolved question is whether the neonatal mouse heart can also regenerate in response to myocardial ischemia, the most common antecedent of heart failure in humans. Here, we induced ischemic myocardial infarction (MI) in 1-d-old mice and found that this results in extensive myocardial necrosis and systolic dysfunction. Remarkably, the neonatal heart mounted a robust regenerative response, through proliferation of preexisting cardiomyocytes, resulting in full functional recovery within 21 d. Moreover, we show that the miR-15 family of microRNAs modulates neonatal heart regeneration through inhibition of postnatal cardiomyocyte proliferation. Finally, we demonstrate that inhibition of the miR-15 family from an early postnatal age until adulthood increases myocyte proliferation in the adult heart and improves left ventricular systolic function after adult MI. We conclude that the neonatal mammalian heart can regenerate after myocardial infarction through proliferation of preexisting cardiomyocytes and that the miR-15 family contributes to postnatal loss of cardiac regenerative capacity.

Modular representations of odorants in the glomerular layer of the rat olfactory bulb and the effects of stimulus concentration.

To study the mechanism whereby odorants are encoded in the nervous system, we studied the glomerular‐layer activity patterns in the rat olfactory bulb evoked by closely related odorants from different chemical families. These odorants had a common straight‐chain hydrocarbon structure, but differed systematically in their functional groups. Neural activity was mapped across the entire glomerular layer by using the [14C]2‐deoxyglucose method. Group responses were averaged and compared by using data matrices. The glomerular activity patterns that resulted from this analysis were comprised of modules. Unique combinations of modules were activated by each odorant, demonstrating what may be part of the neural code for odorants. Most of the modules were clustered together in the bulb, perhaps providing for enhanced contrast between related chemicals by means of lateral inhibition. We also determined whether changes in odorant concentration would affect spatial patterns of glomerular activity. Two odorants, pentanal and 2‐hexanone, evoked different patterns at increased concentrations, with additional glomeruli being recruited at a great distance from glomeruli in which activity was evoked at lower concentrations. Humans report that both of these odorants change in perceived odor with increasing concentration. Three other odorants (pentanoic acid, methyl pentanoate, and pentanol) did not recruit new areas of glomerular activation with increasing concentration, and humans do not report a changed odor across concentrations of these odorants. The results suggest that changes in modular glomerular activity patterns could underlie altered odor perception across odorant concentrations, and they provide additional support for a combinatorial, spatially based code in the olfactory system. J. Comp. Neurol. 422:496–509, 2000.

Chemotopic Odorant Coding in a Mammalian Olfactory System

Systematic mapping studies involving 365 odorant chemicals have shown that glomerular responses in the rat olfactory bulb are organized spatially in patterns that are related to the chemistry of the odorant stimuli. This organization involves the spatial clustering of principal responses to numerous odorants that share key aspects of chemistry such as functional groups, hydrocarbon structural elements, and/or overall molecular properties related to water solubility. In several of the clusters, responses shift progressively in position according to odorant carbon chain length. These response domains appear to be constructed from orderly projections of sensory neurons in the olfactory epithelium and may also involve chromatography across the nasal mucosa. The spatial clustering of glomerular responses may serve to “tune” the principal responses of bulbar projection neurons by way of inhibitory interneuronal networks, allowing the projection neurons to respond to a narrower range of stimuli than their associated sensory neurons. When glomerular activity patterns are viewed relative to the overall level of glomerular activation, the patterns accurately predict the perception of odor quality, thereby supporting the notion that spatial patterns of activity are the key factors underlying that aspect of the olfactory code. A critical analysis suggests that alternative coding mechanisms for odor quality, such as those based on temporal patterns of responses, enjoy little experimental support. J. Comp. Neurol. 503:1–34, 2007.

Multidimensional chemotopic responses to n-aliphatic acid odorants in the rat olfactory bulb

In an effort to understand the means by which similar chemical odorants are encoded in the mammalian brain, we exposed rats to a homologous series of n‐aliphatic acids and mapped the response of the entire olfactory bulb glomerular layer by using a high‐resolution [14C]‐2‐deoxyglucose uptake technique. We found that these similar odorants evoked spatially clustered but distinct responses in the bulb that changed systematically with carbon chain length. In addition to these chemotopic responses, different odorants within the series evoked systematic differences along two other dimensions: amount of deoxyglucose uptake and extent of the glomerular layer showing high activity. Increases along these two dimensions also were correlated with increasing carbon number. The focal glomerular responses were mirrored by responses in deeper bulb layers. Decreasing the odorant concentration decreased the deoxyglucose uptake within focal regions. The focal regions of activity occurred in pairs involving both medial and lateral representations in the bulb, perhaps reflecting the paired medial and lateral projections of olfactory sensory neurons expressing specific types of odorant feature receptor proteins. The observed spatial pattern of response also may explain both the failure of some bulb lesions to interfere with behavioral olfactory responses and the success of other lesions in blocking olfactory responses. These data support a model of parallel, distributed processing of odorants along multiple dimensions. They also support the notion that analyses of the spatial relationships among odorant responses in the olfactory bulb can demonstrate aspects of the mechanism for odor chemical coding. J. Comp. Neurol. 409:529–548, 1999.

Olfactory coding in the mammalian olfactory bulb.

There have been a number of recent approaches to the study of olfactory coding, each of which has its advantages and disadvantages. In the present review, we discuss our own work on this topic, which has involved mapping uptake of [14C]2-deoxyglucose across the entire glomerular layer of the rat main olfactory bulb in response to systematically selected pure odorant molecules. Our strategy to understand the olfactory code has involved four approaches. In the first, we determined whether the system encodes odorants in their entirety, or whether it encodes odorants by representing combinations of molecular features that add together to comprise a neural picture of each odorant. Multiple odorant features appeared to be coded by multiple receptors. Our second strategy examined the ways that such features are represented. We stimulated rats with odorants that differed greatly in their molecular structure to be able to identify a set of odorant feature response domains. Our third approach asked how odorants with very small differences in molecular structure are coded, and we found systematic differences in the representation of such features within response domains. Finally, we were able to predict odor perception from the neural representations of odorants that differed in only a single aspect of their structure. Using these strategies, we have been able to learn some of the rules by which the olfactory code operates. These rules have allowed us to predict where previously unmapped molecules would be represented and how differences in molecular representations affect olfactory perceptions.

Spontaneous versus Reinforced Olfactory Discriminations

When the major response domains in the rat olfactory bulb that are evoked by odorant enantiomers are compared, some of these odorant pairs do not show significantly different activity patterns. Such pairs are not spontaneously discriminated in a behavioral test. We show here that even these similar odorants appear to evoke different activity patterns when every data point in a glomerular activity array is compared. These odorants also can be discriminated if they are subjected to differential reinforcement. These data suggest that the method chosen to assess olfactory discrimination will reveal different olfactory capabilities of rats. The small differences in glomerular activity that probably exist between any pair of odorants may serve as a basis for odor discrimination when rats are differentially reinforced, thereby establishing the remarkable limits of rat olfactory perception. At the same time, the major differences in glomerular responses appear to serve as the normal basis for spontaneous odor discrimination.

Relational representation in the olfactory system

The perceptual quality of odors usually is robust to variability in concentration. However, maps of neural activation across the olfactory bulb glomerular layer are not stable in this respect; rather, glomerular odor representations both broaden and intensify as odorant concentrations are increased. The relative levels of activation among glomeruli, in contrast, remain relatively stable across concentrations, suggesting that the representation of odor quality may rely on these relational activity patterns. However, the neural normalization mechanisms enabling extraction of such relational representations are unclear. Using glomerular imaging activity profiles from the rat olfactory bulb together with computational modeling, we here show that (i) global normalization preserves concentration-independent odor-quality information; (ii) perceptual similarities, as assessed behaviorally, are better predicted by normalized than by raw bulbar activity profiles; and (iii) a recurrent excitatory circuit recently described in the olfactory bulb is capable of performing such normalization. We show that global feed-forward normalization in a sensory system is behaviorally relevant, and that a center-surround neural architecture does not necessarily imply center-surround function.

Odorant molecular length: One aspect of the olfactory code

Organic acid odorants of differing carbon number produce systematically different spatial patterns of [14C]2‐deoxyglucose uptake in the glomerular layer of the olfactory bulb. Because increasing carbon number correlates with progressive increases in several molecular features, including hydrophobicity, length, and volume, we determined which of these properties was most associated with systematic changes in the location of an anterior, dorsomedial module responding to fatty acids. We exposed groups of rats to two series of organic acids that each had the same number of carbons, but differed in their hydrocarbon structures. These straight‐chained, branched, cyclic, and double‐bonded molecules differed independently in hydrophobicity, length, and volume. The only molecular property that was strongly correlated with the location of the module was molecular length, suggesting that this molecular feature is the principal determinant of the chemotopic organization of glomeruli within the module. We also found that distinct hydrocarbon structures produced large differences in spatial patterns of 2‐deoxyglucose uptake in posterior parts of the bulb. Even subtly distinct structural isomers evoked posterior responses that differed greatly. The odorant 2‐methylbutyric acid evoked much greater uptake in the posterior bulb than did its structural isomer 3‐methylbutyric acid (isovaleric acid). These data suggest that posterior portions of the bulb may encode specific steric features of odorant molecules and that some odorant features may have an inherent or acquired greater representation than do others. J. Comp. Neurol. 426:330–338, 2000.

Functional Mapping of the Rat Olfactory Bulb Using Diverse Odorants Reveals Modular Responses to Functional Groups and Hydrocarbon Structural Features

In an effort to understand the olfactory code of rats, we collected more than 1,500,000 measurements of glomerular activity in response to 54 odorants selected to provide differences in functional groups and hydrocarbon structure. Each odorant evoked a unique response pattern by differentially stimulating clusters of glomeruli, called modules. Odorants sharing specific aspects of their structure activated the same modules, allowing us to relate responses to structure across approximately 80% of the glomerular layer. The most obvious relationship was between the presence of particular oxygen‐containing functional groups and the activity of glomeruli within dorsal modules. Functional group‐specific responses were observed for odorants possessing a wide range of hydrocarbon structure, including aliphatic, cyclic, and aromatic features. Even formic acid and acetone, the simplest odorants possessing acid or ketone functional groups, respectively, stimulated modules specific for these functional groups. At the same time, quantitative analysis of pattern similarities revealed relationships in activation patterns between odorants of similar hydrocarbon structure. The odorant responses were reliable enough to allow us to predict accurately specific aspects of odorant molecular structure from the evoked glomerular activity pattern, as well as predicting the location of glomerular activity evoked by novel odorants. J. Comp. Neurol. 449:180–194, 2002.

A learned odor evokes an enhanced Fos-like glomerular response in the olfactory bulb of young rats

Young rats exposed to peppermint odor and reinforcing tactile stimulation from postnatal days (PND) 1-18 increase their preference for that odor relative to controls. This early olfactory memory is accompanied by an 80% increase in the density of glomerular-layer cells displaying Fos-like immunoreactivity in response to the learned odor on PND 19. The difference is observed in midlateral portions of the olfactory bulb that align with foci of 2-deoxyglucose (2-DG) uptake in adjacent sections. Trained and control animals are not different in the Fos-like response of juxtaglomerular cells within ventrolateral 2-DG foci. Ratios of midlateral/ventrolateral response differ significantly between trained and control animals and include differences among cells of three staining intensities. These ratios are correlated with ratios of 2-DG uptake (midlateral/ventrolateral foci), which also differ significantly between trained and control rats. Juxtaglomerular cells associated with 2-DG foci also express Egr-1-like immunoreactivity. However, the midlateral Egr-1 response does not differ between trained and control rats. These results show that early memories can be associated with an increased Fos-like response in a primary sensory area of the CNS. They also suggest that only specific regions within the olfactory bulb are modified following the learning of a given odor in early life.