Retrieval of developmental social memories requires activation of the adult-born granule cell-CA2 circuit in mice

Reviewer #1 (Public Review):

Summary:
This manuscript provides some valuable findings concerning the hippocampal circuitry and the potential role of adult-born granule cells in an interesting long-term social memory retrieval. The behavior experiments and strategy employed to understand how adult-born granule cells contribute to long-term social discrimination memory are interesting.

I have a few concerns, however with the strength of the evidence presented for some of the experiments. The data presented and the method described is incomplete in describing the connection between cell types in CA2 and the projections from abGCs. Likewise, I worry about the interpretation of the data in Figures 1 and 2 given the employed methodology. I think that the interpretation should be broadened. This second concern does not impact the interest and significance of the findings.

Strengths:
The behavior experiments are beautifully designed and executed. The experimental strategy is interesting.

Weaknesses:
The interpretation of the results may not be justified given the methods and details provided.

Reviewer #2 (Public Review):

Summary:
Laham et al. investigate how the projection from adult-born granule cells into CA2 affects the retrieval of social memories at various developmental points. They use chemogenetic manipulations and electrophysiological recordings to test how this projection affects hippocampal network properties during behavior. I find the study to be very interesting, the results are important for our understanding of how social memories of different natures (remote or immediate) are encoded and supported by the hippocampal circuitry. I have some points that I added below that I think could help clarify the conclusions:
- My major concern with the manuscript was that making the transitions between the different experiments for each result section is not very smooth. Maybe they can discuss a bit in a summary conclusion sentence at the end of each result section why the next set of experiments is the most logical step.
- In line 113, the authors say that "the DG is known to influence hippocampal theta-gamma coupling and SWRs". Another recent study Fernandez-Ruiz et al. 2021, examined how various gamma frequencies in the dentate gyrus modulate hippocampal dynamics.
- Having no single cells in the electrophysiological recordings makes it difficult to interpret the ephys part. Perhaps having a discussion on this would help interpret the results. If more SWRs are produced from the CA2 region (perhaps aided by projections from abGC), more CA2 cells that respond to social stimuli (Oliva et al. 2020) would reactivate the memories, therefore making them consolidate faster/stronger. On the other hand, the projections from abGC that the authors see, also target a great deal of PV+ interneurons, which have been shown to pace the SWRs frequency (Stark et al 2014, Gan et al 2017), which further suggests that this projection could be involved in SWRs modulation.
- The authors should cite and discuss Shuo et al., 2022 (A hypothalamic novelty signal modulates hippocampal memory).
- I think the authors forgot to refer to Fig 3a-f, maybe around lines 163-168.
- Are the SWRs counted only during interaction time or throughout the whole behavior session for each condition?
- Figure 3t shows a shift in the preferred gamma phase within theta cycles as a result of abGC projections to CA2 ablation with CNO, especially during Mother CNO condition. I think this result is worth mentioning in the text.
- Figure 3u in the legend mention "scale bars = 200um", what does this refer to?
- What exactly is calculated as SWR average integral? Is it a cumulative rate? Please clarify.
- Alexander et al 2017, "CA2 neuronal activity controls hippocampal oscillations and social behavior", examined some of the CA2 effects in the hippocampal network after CNO silencing, and the authors should cite it.

Strengths:
Behavioral experiments after abGC projections to CA2 are compelling as they show clearly distinct behavioral readout.

Weaknesses:
Electrophysiological experiments are difficult to interpret without additional quantifications (single-cell responses during interactions etc.)

Reviewer #3 (Public Review):

Laham et al. present a manuscript investigating the function of adult-born granule cells (abGCs) projecting to the CA2 region of the hippocampus during social memory. It should be noted that no function for the general DG to CA2 projection has been proposed yet. The authors use targeted ablation, chemogenetic silencing, and in vivo ephys to demonstrate that the abGCs to CA2 projection is necessary for the retrieval of remote social memories such as the memory of one's mother. They also use in vivo ephys to show that abGCs are necessary for differential CA2 network activity, including theta-gamma coupling and sharp wave-ripples, in response to novel versus familiar social stimuli.

The question investigated is important since the function of DG to CA2 projection remained elusive a decade after its discovery. Overall, the results are interesting but focused on the social memory of the mother, and their description in the manuscript and figures is too cursory. For example, raw interaction times must be shown before their difference. The assumption that mice exhibit social preference between familiar or novel individuals such as mother and non-mother based on social memory formation, consolidation, and retrieval should be better explained throughout the manuscript. Thus, when describing the results, the authors should comment on changes in preference and how this can be interpreted as a change in social memory retrieval. Several critical experimental details such as the total time of presentation to the mother and non-mother stimulus mice are also lacking in the manuscript. The in vivo e-phys results are interesting as well but even more succinct with no proposed mechanism as to how abGCs could regulate SWR and PAC in CA2.

The manuscript is well-written with the appropriate references. The choice of the behavioral test is somewhat debatable, however. It is surprising that the authors chose to use a direct presentation test (presentation of the mother and non-mother in alternation) instead of the classical 3-chamber test which is particularly appropriate to investigate social preference. Since the authors focused exclusively on this preference, the 3-chamber test would have been more adequate in my opinion. It would greatly strengthen the results if the authors could repeat a key experiment from their investigation using such a test. In addition, the authors only impaired the mother's memory. An additional experiment showing that disruption of the abGCs to CA2 circuit impairs social memory retrieval would allow us to generalize the findings to social memories in general. As the manuscript stands, the authors can only conclude the importance of this circuit for the memory of the mother. Developmental memory implies the memory of familiar kin as well.

The in vivo ephys section (Figure 3) is interesting but even more minimalistic and it is unclear how abGCs projection to CA2 can contribute to SWR and theta-gamma PAC. In Figure 1, the authors suggest that abGCs project preferentially to PV+ neurons in CA2. At a minimum, the authors should discuss how the abGCs to PV+ neurons to CA2 pyramidal neurons circuit can facilitate SWR and theta-gamma PAC.

Finally, proposing a function for 4-6-week-old abGCs projecting to CA2 begs two questions: What are abGCs doing once they mature further, and more generally, what is the function of the DG to CA2 projection? It would be interesting for the authors to comment on these questions in the discussion.